Process for extracting DNA from biological samples

A biological sample and chip technology, which is applied in biochemical equipment and methods, microbial determination/inspection, sugar derivatives, etc. The effect of less reagent consumption, enhanced safety, and simple chip fabrication

Inactive Publication Date: 2005-07-06
SHANGHAI INST OF MICROSYSTEM & INFORMATION TECH CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the currently reported chips are mainly based on the traditional silicon micromachining process. The cost of silicon processing chips is still high, and the processing technology is complicated, which is not conducive to the reduction of detection costs.
[0008] In recent years, microchips made of plastic materials have attracted people's attention due to their low cost and convenient processing. However, due to the limitations of their own material properties, plastic materials cannot be used as solid supports to complete the extraction of DNA.
Therefore, the sample DNA extraction chip made of plastic material has not been reported.

Method used

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  • Process for extracting DNA from biological samples
  • Process for extracting DNA from biological samples
  • Process for extracting DNA from biological samples

Examples

Experimental program
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Effect test

preparation example Construction

[0030] see figure 1 , the preparation method of polydimethylsiloxane chip, comprises the steps:

[0031] (1) Preparation of the mould: transfer the designed chip structure to a mask 1 that adopts a silicon wafer or a glass wafer as a base material and is coated with a photoresist, then photoetches 4, develops 5 by a conventional method, and obtains A mold provided with protruding runners 2;

[0032] Said photoresist is a conventional material used in photolithography, such as SU8 negative photoresist; (SU-8, 2025, Microchem).

[0033](2) Preparation of the chip: place the small glass column 3 on the top of the protruding runner 2 of the mold, then cast the mixture of polydimethylsiloxane and curing agent on the mold 6, solidify, and demould 7, that is Obtain a polydimethylsiloxane sheet with micro-channels, bond and package with another polydimethylsiloxane sheet or glass sheet, and finally form a chip made of polydimethylsiloxane ,Such as figure 2 , the channel formed by...

Embodiment 1

[0035] The DNA in the Escherichia coli culture solution is extracted, and the steps are as follows:

[0036] 1) Introduce 5ul micro-magnetic bead dilution into the chip, place a magnet under the channel to absorb the magnetic beads, pressurize the liquid after three minutes of adsorption to discharge the liquid from the chip, and fix the magnetic beads in the chip through the adsorption of the magnet, such as image 3 ;

[0037] 2) Escherichia coli Ecoli DH5α was inoculated and cultured overnight on a shaking table.

[0038] 3) Take away the magnet, add 20 μL of the bacterial solution to 50 μL of adsorption buffer (4% Triton X-100, 0.5M NaCl, 20% PEG 8000), slowly introduce into the chip, mix and then introduce into the chip, and place it at 37°C 15 minutes, then place a magnet under the channel to absorb the magnetic beads, pressurize at the same time to discharge the liquid at a flow rate of 2 μL / min, collect the recovered liquid, and obtain the sample 1

[0039] 4) A magn...

Embodiment 2

[0046] The DNA in the PCR product is extracted, and the steps are as follows:

[0047] 1) Introduce 5ul micro-magnetic bead dilution into the chip, place a magnet under the channel to absorb the magnetic beads, pressurize the liquid after three minutes of adsorption to discharge the liquid from the chip, and fix the magnetic beads in the chip through the adsorption of the magnet, such as image 3 ;

[0048] 2) Take away the magnet, add 10 μL of the PCR product to 20 μL of adsorption buffer and slowly introduce it into the microchannel of the chip, mix it, and place it at 37 ° C for 10 minutes, then place a magnet under the microchannel to absorb the magnetic beads, and pressurize at the same time The liquid is discharged so that the flow rate is 2 μL / min;

[0049] 3) A magnet is still placed under the chip, and 20ul of 70% ethanol is introduced into the microchannel for flow cleaning;

[0050] 4) Place the chip in a 37°C incubator for 2 minutes to dry the magnetic beads;

...

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Abstract

This invention relates to a biological sample DNA extraction method, which comprises the following steps: a, leading the micro magnetic leads with carboxyl mark into the micro channel of the chip and absorbing and fixing it inside the channel to form the micro flow chip of the sample; b, mixing the sample and DNA absorbing liquid and then leading into the chip and mixing it with the micro magnetic leads, wherein the DNA in sample is absorbed on the leads with buffer liquid flowing in for washing and then collecting the washing liquid to get the high purified DNA.

Description

technical field [0001] The invention relates to a method for extracting DNA in biological samples, in particular to a method for extracting DNA in biological samples by using micro magnetic beads in a microfluidic chip. Background technique [0002] When analyzing DNA in biological samples, steps such as DNA extraction, DNA amplification (PCR amplification) and amplification detection (electrophoresis detection or hybridization detection) are often required. At present, the routine DNA extraction methods in the laboratory mainly adopt alkali cleavage method or phenol-chloroform method, that is, liquid-liquid methods (Liquid-liquid Methods), which require centrifuges and other equipment, and the operation steps are time-consuming and tedious. Phenol and chloroform are also harmful to the human body. Toxic. [0003] The newer method is to use the SPE (solid phase extraction) method, that is, the solid phase extraction method. Compared with traditional liquid-liquid methods, ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07H21/04C12Q1/68
Inventor 杨梦苏陈翔赵建龙王斌
Owner SHANGHAI INST OF MICROSYSTEM & INFORMATION TECH CHINESE ACAD OF SCI
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